It is our goal to define precisely the chemical nature of the immuno-reactive Thomsen-Friedenreich (T)-specific structures which are characteristically associated with human adenocarcinomata and to which the adenocarcinoma patient shows impressive cellular as well as humoral immune reactions. These immunogenic structures are not present in reactive form in comparable benign structures and not as far as we have found in melanoma and tumors of the nervous system. The active substances will be compared with corresponding structures on epithelial tissues of healthy persons and those with benign diseases. On these latter tissues T specificity is masked by a terminal sugar, generally sialic acid. We want to establish fully by standard chemical, immunochemical, and physical procedures nature and membrane localization of these T-specific reactive structures in adenocarcinoma cells, 1) from tissues obtained freshly at surgery and 2) from those grown in tissue culture. We will also study tissue culture supernatants since breast carcinoma cells have been found to shed T-, Tn- and M- & N-like substances into their environment. Our efforts will be concentrated on primary human breast carcinomata and on their metastatic lesions. We will extract from these tissues glycoprotein and glycolipid fractions. From these we will isolate T- & Tn- and M- & N-Specific glycopeptides and glycolipids. We will cleave the glycosidically linked oligosaccharide chains by beta-elimination with NaB(3H)4 from peptide structures or by Os-catalyzed IO4- oxidation from the glycolipids. We will determine what part the peptide and lipid carriers of the T-specific structures play in the host's immune response and if we can pinpoint specific areas in the peptide chain responsible for this.